Intein을 이용한 대장균에서의 Trichoderma reesei 유래의 Cellobiohydrolase I 섬유소 결합 도메인의 발현

• 영문명: Intein-mediated expression of Trichoderma reesei Cellobiohydrolase I Cellulose Binding Domain in E. coli
• 발행기관: 강원대학교 산업기술연구소
• 저자명: 최신건(Choi, Shin-Geon)
• 간행물 정보: 『산업기술연구』vol.36, 33~38쪽, 전체 6쪽
• 주제분류: 공학 > 공학일반
• 파일형태: PDF
• 발행일자: 2016.09.30

영문 초록

Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli . Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.

목차

1. INTRODUCTION

2. MATERIALS AND METHODS

3. RESULTS AND DISCUSSION

4. CONCLUSION

키워드

Trichoderma reesei cellulose binding domain intein 재조합발현(recombinant expression)